Preparation and properties of creatine kinase from the breast muscle of normal and dystrophic chicken (Gallus domesticus)

Abstract

1. The purification of creatine kinase from normal and genetically dystrophic chicken breast muscle is described. Enzyme recovery was significantly lower from dystrophic muscle. 2. Both enzymes had the same number of reactive and total thiol groups and had similar specific activities and similar amino acid compositions. 3. No significant differences were observed in sedimentation, electrophoretic or kinetic properties. 4. Peptide ;maps' showed no significant differences, and electrophoresis of partial acid hydrolysates of the labelled enzymes suggested that corresponding amino acid sequences around all the thiol groups were very similar. 5. The enzymes showed identical temperature stabilities. 6. No significant differences between the enzymes from normal and dystrophic muscle were observed.

The amino acid sequence of the peptide containing the thiol group of creatine kinase from normal and dystrophic chicken breast muscle. Comparison of some of the immunological properties of the antibodies developed in rabbits against these enzymes

“…The crossreaction between normal muscle creatine kinase and antisera against the dystrophic muscle enzyme (or vice versa) observed by immunodiffusion and by immunoelectrophoretic experiments further suggests that the enzymes from normal and dystrophic chicken muscle are similar in structure. The results of the present study, the identical amino acid sequence of the peptides containing the reactive thiol group from both the normal and dystrophic chicken muscle enzymes and the immunological similarities of the two enzymes are in accord with the similarity of the two enzymes observed by Roy et al (1970).…”

“…Further, the replacement by a glutamic acid residue of cysteine would require at least two (and more likely three) nucleotide base changes, which seems rather unlikely to occur (Morgan et al, 1966). No significant differences were observed in the Km values for normal and dystrophic chicken (Roy et al, 1970) or mouse (Hooton & Watts, 1966a) enzymes. However, only 50% inactivation with dystrophic mouse enzyme (Roy et al, 1970) was observed when their free thiol groups were blocked with iodoacetate, compared with 100% inactivation for the normal enzymes of both species.…”

“…It was decided to investigate this problem in further detail by using New Hampshire chickens with hereditary myopathy. The earlier part of this work has been presented (Roy et al, 1969(Roy et al, , 1970Roy, 1971Roy, , 1972 and no significant differences in creatine kinases from normal and dystrophic chicken muscle were observed by various criteria. The present paper is concerned with the amino acid sequence around the reactive thiol groups of normal and dystrophic muscle creatine kinase and serological comparison of the two enzymes by immunodiffusion, immunoelectrophoretic and interfacial tests.…”

“…With bovine brain enzyme, Atherton et al (1970) also did not find complete inhibition under certain conditions of enzyme concentration and temperature. Roy et al (1970) suggested that if the thiol groups in the two subunits were not identical and if the reactivity of one of them was modified by subunit interaction, the incompleteness of the inhibition of the chicken enzyme could be explained, and this would also be applicable to the mouse enzyme.…”

A Protein That Binds Specifically to the M-Line of Skeletal Muscle Is Identified as the Muscle Form of Creatine Kinase

“…It should be noted that the scheme for purification of M-line protein of Morimoto and Harrington (11) is a considerably simpler method for isolation of MM-creatine kinase than those used previously for isolating the chicken enzyme (13,20). It is possible that one or more of the procedures will be useful in preparing MM-creatine kinase from other sources, or in achieving a simpler purification method for the BB or MB enzyme (14).…”